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Cells and Model Organisms

Beginners Guide to Setting Up Migration and Invasion Assays

So you have a gene or protein that you think may be involved in migration or invasion and the next step is to embark on migration assays. These assays are useful for testing fundamental migratory processes, such as embryonic development, immune response, metastasis and angiogenesis. For a long time these have been an invaluable mainstay…

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Cell Culture is No Longer Flat: Three Dimensional Cell Culture

Three dimensional cell culture mimics the extracellular matrix (ECM) that offers the structure and support for cells in vivo, thus creating the complex architecture and network required for cellular communication. For 3D cell culture beginners (or enthusiasts), the information available may seem overwhelming. It sure was for me. But it can be simplified. For example,…

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How to Culture Primary Human Bronchial Epithelial Cells

Human bronchial epithelial cells (HBECs) are a challenge to culture. As highly specialised cells that exist in carefully ordered multi-layered structures, they are especially fickle and finding optimum conditions to keep them happy is tricky. The cultures are also extremely sensitive to tiny changes in routine or environment. However, there are certain basic principles that…

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Troubleshooting and Optimizing your Mouse Breedings

Anyone working with laboratory animals has probably realized that simply putting two animals together does not always yield new offspring and reliable continuity of the animal line – unfortunately animal husbandry isn’t that simple! Of course, apart from making sure that the two animals put together are from different genders, there are a lot of…

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Measuring Invasion – Round is the New Flat

I’m sure many of us are aware that the world of cancer research is exploding with the idea of cancer stem cells. This exciting hypothesis suggests that there is a small population of cells in the tumor that have stem cell-like properties. These stem-like cells are able to proliferate and differentiate into all the different…

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Practical Secrets of the Bacterial World for the Uninitiated

As a protein biochemist where bacteria were mere workhorses, imagine my surprise when I began work in a bonafide micro lab! I discovered that bacteria could be much fussier than my good ol’ cloning and expression friends E. coli DH5ɑ or BL21. One broth would not do for all, some even required blood! No, no,…

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Five Great Resources to Learn About Stem Cells

Whether you’re already in the field or an undergrad looking to enter the scene, here are some great places to keep up to date with the latest news and trends in stem cells. Listen About It For auditory learners, or people that listen to music on their way to the lab, you could switch it…

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Furthering Insight and Productivity in Cell Biology with Real-Time Quantitative Live-Cell Analysis

In this webinar you’ll learn how live-cell analysis offers you a whole range of new tools for studying your cells in culture, in real-time. The main points we’ll cover are: How live-cell imaging can allow you to perform detailed, real-time analyses, such as reporter assays, live phenotypic analysis, cell migration, and more, on your cultured cells, without taking them…

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Isolation and Culture of Primary Cells from Bone Marrow

Bone marrow stromal cells have been one of the more intensively studied adult stem cell types over the past 50 years. Great therapeutic interest in this adult stem cell type has been based on its complex and diverse biological properties including their ability to support a hematopoietic environment, possess anti-inflammatory qualities, and differentiate down multiple…

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How to Become Immortal: Generation of Immortal Cell Lines

Normal cells are unable to replicate past several rounds of proliferation (termed the Hayflick limit) as with each round of proliferation the telomeres shorten. When the telomeres reach a critically reduced length, DNA damage is triggered leading to cellular senescence. Therefore, if you tried to culture a primary cell population it would eventually die unless…

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Culturing Dendritic Cells – the Factors that Matter

Murine bone marrow derived dendritic cells (BMDCs) are one of the easiest primary cell cultures to generate. The beauty lies in the fact that in the end you have a large quantity of robust DCs that can be matured and used to study a variety of DC functions and DC-T-cell interactions. But there are a…

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A Beginner’s Guide to Culturing Mouse Embryonic Stem Cells

There is something undeniably special about embryonic stem cells (ESCs) and not just because they can produce every cell type in the adult body. In vivo, ESCs are a transitory state of early development, which has been captured indefinitely in vitro. Whether you are a hardened cell culture enthusiast or have just graduated from the…

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Heat Inactivation of Serum for Tissue Culture – Is it Necessary?

In the cell culture practice, heat inactivation of serum products has always been accepted and is one of the basic protocols passed on to new cell culturists. There is no strict standard protocol for heat inactivation, some say incubate at 56 °C for 30 minutes, while some say it can be efficiently performed using temperatures…

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Top Tips on How to Prevent Cell Line Cross-Contamination

Recently we wrote an article about widespread cell culture contamination and how to detect it. This follow-up article will provide practical tips on avoiding cross-contamination in the first place. Be Cautious While Working The first way of cross-contaminating cultures is by accidentally mixing two cultures together, which may lead to an unintended co-culture or the displacement…

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Cell Culture: a Case of Mistaken Identity

While working in a UK university, I met a researcher who loved Italy much more than the UK. I asked her why she had left her favourite country. She told me that before coming to the UK, she had a 2-year fellowship in Italy where she was getting some promising results and had the chance…

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12 Top Tips for Working in Your Cell Culture Hood

Whether you’re about to become keeper of the cells, or are just passing through to run a pilot study, knowing how to use the biosafety cabinet is just as essential as knowing how to use the fume hood when working with non-crawling, chemical reagents. We’ve seen a brief protocol for how to use the biosafety…

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Six Ways to Measure T Cell Responses

T cells can be problematic to characterise because they have a wide variety of subtypes and because of the technical difficulties of studying the membrane-bound T cell receptor, but there are situations where you want to be able to do this such as analysing the degree to which immunological memory has been induced to measuring…

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Which Cytokine Will I Get? How to Stimulate Human Cytokine-Producing Cells

Cells are like people: depending on their current environment, past experiences and their genetic make-up they will react differently. Treat cells in different ways, and they will produce different cytokines. There are a lot of protocols out there for stimulating cells. Depending on the species of cells you plan to stimulate, different protocols are available…

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Tips for a Happily Functioning Tissue Culture Room

You walk into your tissue culture room to find a window open, the incubator’s humidity tray empty and a pipette lying on its side in the hood on a used glove… After you have found and torn to shreds the person responsible for this monstrous act (!!), consider posting the below tips to help those…

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Microbiology 101 by Aunt Yersinia

We are pleased to announce that the famous maid of microbiology, dear old Aunt Yersinia, has agreed to start writing a microbiology and molecular biology advice column for Bitesize Bio. She will be free to answer your most pressing questions sent to:  auntyersinia@bitesizebio.com By way of introduction, Aunt Yersinia is bestowing 8 spores of knowledge garnered…

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Getting in Deep: How to Deep Clean a Tissue Culture Hood

One of the most exciting aspects of being a biologist is getting opportunities to examine how and why living organisms behave the way they do. We have technology that enables us to obtain images at sub-cellular levels, and the skills to work directly with the micro-environments essential for the progression of life. However, at the…

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Seven Things That Really Annoy Me About Tissue Culture

The tissue culture facility can be one of the most important places in the lab.  Many researchers spend hours in the hoods isolating primary cell lines and tissue, generating samples for western blot analysis, ELISA…  the list goes on. It is also one of the places in which scientists tend to butt heads. An improperly…

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Mycoplasma: The Hidden Anarchist of Cell Culture

It is the black death of cell culture. Scientists don’t dare utter its name and many a graduate student has fallen victim to its indiscriminate menace. These stealthy anarchists infiltrate quietly but deliberately until their numbers swell and then they attack in strength, overwhelming their victims before they can put up a fight! What is…

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What is Sterile? Find Your Way around a Sterile Tissue Culture Hood

You’ve been told that maintaining a sterile environment in a tissue culture hood is vital to preventing contamination of cell cultures. But what exactly is meant by sterile? The definition of sterile is ‘completely clean, sanitized, and free of all forms of life’. Obviously you still want your cells and/or any other organisms you are…

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I can’t breathe: Is 20% Oxygen Always the Appropriate Level for Cell Culture?

Researchers spend considerable time and money on proper experimental design for in vitro cell culture, so why is it so difficult for cells in culture to have the same physiological function as in our body? You can be working in a sterile environment, utilizing high end laboratory cell culture equipment and have supreme cell culture…

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What’s in a Number: Getting the Right Passage in Cell Culture

Getting the Right Passage Number Using an American Type Culture Collection (ATCC) reference strain for every experiment would be great, but not all that practical. So, most labs subculture their cells into a new vessel. This subculture is also known as a “passage.” A passage number is the number of times a cell culture has…

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Outgrown the Roost: Passaging Suspension Cells

Previously, you have learned about passaging adherent cells and read a quick protocol to make it happen. In this article, I will talk about passaging suspension cells. Some cells naturally live in suspension in body fluids and do not attach to surfaces, such as cells of hematopoietic origin found in our bloodstream. Culturing these suspension…

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Outgrown the Roost: A Quick Protocol for Passaging Adherent Cells

Your cells have dutifully “sat down”, stuck to the bottom of the dish and replicated, replicated, replicated. You see a swarm of cells that have reached confluency and filled the entire plate. It is time to split them. This can be a relatively easy ritual, but missteps can lead to dead floating cells and the…

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Outgrown the Roost: Passaging Adherent Cells, the Basic Process

Splitting, passaging, subculturing… whatever you call it, while the specifics of passaging adherent cells will depend upon the individual cell type, the basic process involves four simple steps. These basic steps are outlined below. Rinse Cells With a Balanced Salt Solution (BSS) Prior to detaching cells from the dish, it is important to aspirate off…

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Principles and Mechanisms of Mammalian Cell Transfection

Mammalian cell transfection is a technique commonly used to express exogenous DNA or RNA in a host cell line (for example, for generating RNAi probes). There are many different ways to transfect mammalian cells, depending on the cell line characteristics, desired effect, and downstream applications. In this article, I will review the different methods of…

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Zero Tolerance: A Perfectionist’s Guide to Aseptic Technique

Arguably, molecular biology is impossible without microbiology – even if you work exclusively with transgenic mice, you may one day need to amplify a vector in E. coli. And microbiology is definitely impossible without good aseptic technique. The main principle of good microbiological practice is a zero tolerance approach: it’s good to be a little…

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How Pure is Your Cell Culture Broth? Comparing Mycoplasma Detection Kits

Mycoplasmas are the most difficult-to-detect organisms in your eukaryotic cell culture. And they can be the most dangerous; they can disrupt cell growth and differentiation and even apoptotic patterns without you even knowing what’s going on until it’s too late. Traditional cell culture methods can take up to a month to yield results, which means…

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A Beginner’s Guide to Storing Biological Materials

In a typical biology lab, you may encounter many types of biological materials, including cells, bodily fluids, purified DNA and RNA, enzymes, bacterial cultures, body parts, and whole animals. In order to perform experiments that yield quality results, samples need to be stored properly in order to preserve their activity or integrity. Beginning students and…

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Got Phage? Here’s how to get rid of it.

Summertime… The birds are singing, the trees are growing. Your tissue culture has sprouted yeast contamination, your yeast culture is happily growing bacteria. Your bacterial culture was growing calmly and predictably, dividing every twenty minutes, but suddenly its optical density has dropped, and it’s full of some sort of filaments and clumps. Or you did…

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3 Ways to Abuse Biological Safety Cabinets

As we learned in my previous article, cell culture hoods have many names. As if that wasn’t enough, they are all-too-often misunderstood and mistreated, which can lead to dangerous situations harmful for both the worker and the general lab environment. Here are three common ways that workers abuse biological safety cabinets; make sure you don’t…

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Biological Safety Cabinets and Culture Hoods: Know The Difference

Biological safety cabinets, laminar flow hoods, clean hoods and culture hoods are all common names for those essential pieces of equipment that you use in cell culturing. The terms are used inter-changeably, but in fact there are lots of different types of culture hoods, each of which does a different job. Knowing which is which…

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